睫状神经营养因子基因转染延缓失神经肌萎缩的作用

背景睫状神经营养因子(CNTF)对失神经支配骨骼肌萎缩有防治作用,但是否可以应用CNTF基因转染方法延缓失神经支配骨骼肌萎缩尚不清楚.目的探索一次性电穿孔介导CNTF转染延缓失神经骨骼肌萎缩的疗效.设计随机对照研究.地点和对象在上海交通大学附属第一人民医院动物房完成,雄性SD大鼠36只,年龄两三个月,体质量250～300g,由上海中科院实验动物中心提供.干预制备36只大鼠右下肢腓肠肌失神经支配模型,随机平均分成失神经对照组,CNTF基因转染组.两组于术后2,4,8周分别取6只大鼠进行实验评价.主要观察指标两组术后2,4,8周肌湿重维持率、肌细胞截面积、肌肉蛋白含量、胶原纤维与肌细胞面积比和细胞凋亡数.结果2周和4周CNTF基因转染组的肌湿重维持率、肌细胞截面积和肌肉蛋白含量均明显高于失神经对照组(P＜0.05),而胶原纤维与肌细胞面积比和细胞凋亡数明显低于失神经对照组(P＜0.05).8周后,两组间各参数无明显区别(P＞0.05).结论一次性电穿孔介导CNTF基因转染可延缓失神经骨骼肌萎缩4周.

Effects of gene transfer of ciliary neurotrophic factor on delaying of denervated muscular atrophy

BACKGROUND: Ciliary neurotrophic factor(CNTF) was proved to be effective on delaying the atrophy of denervated skeletal muscles. However, it is still unclear whether the gene transfer of CNTF can delay the atrophy of denervated skeletal muscle.OBJECTIVE: To explore the therapeutic effect of one-time gene eletrotransfer of CNTF on delaying denervated muscle atrophy.DESIGN: A randomized controlled trial.SETTING and PARTICIPANTS: The research was done at the Laboratory for Animals, Shanghai First People' s Hospital, Shanghai Jiao Tong University. Twenty-six male SD rats, aged 2 - 3 months, weighing 250 - 300 g were used(provided by Shanghai Laboratory Animal Center, Chinese Academy of Sciences).INTERVENTIONS: A model of denervated gastrocnemius of right lower limb was established in thirty-six rats. They were divided into two groups: denervation control group and CNTF gene transfer group. Experimental evaluation was performed for six rats of each group 2, 4 and 8 weeks after operation.MAIN OUTCOME MEASURES: Rate of wet muscle mass preservation,muscle cell cross-section area, protein amount, area rate of the collagen fibers versus muscle cells and the number of apoptotic muscle cells were measured at 2, 4 and 8 weeks.RESULTS: On the 2nd and 4th week, the rate of wet muscle mass preservation, the muscle cell cross-section area and the protein amount were significantly greater in CNTF gene transfer group than those in denervation control group ( P ＜ 0.05 ), while the area rate of the collagen fibers versus muscle cells and the number of apoptotic muscle cells were significantly fewer in CNTF gene transfer group than those in denervatian control group( P ＜ 0. 05) . After 8 weeks, there was no significant difference between two groups in regard to the parameters mentioned above ( P ＞ 0. 05 ).CONCLUSION: One-time gene transfer of CNTF mediated by electroporation could attenuate the denervated muscle atrophy for 4 weeks.